The use of cassava as foodstuffs has been widely developed. Modifying it into a mocaf (modified cassava flour) is one of the cassava utilization. This research aimed to process cassavas into mocaf utilizing lactic acid bacteria and to obtain a better mocaf in terms of its physicochemical parameters. Bacteria used were L. plantarum, L. fermentum, and L. paracasei, which can ferment cassava to mocaf. The fermentation process was carried out by two fermentation duration of 48 hours and 72 hours, followed by draining and drying using the oven at 50 °C for 6 hours. This research analyzed mocaf's physicochemical properties such as water content, fat content, protein content, ash content, carbohydrate content, whiteness, and acidity. A Factorial Randomized Block Design with two replications was applied as the research design. If the test result showed that the tested sample has a significant difference at the level of significance of 0.05, it then subjects to the further Duncan test, using SPSS. The result showed that the use of L. paracasei produced best characteristics mocaf with a high protein content of 1.44%, an ash content of 0.31%, a white degree of 102.20, and a low degree of acid of 3.66.

Abstract. Hartati, Fathoni A, Kurniawati S, Hartati NS, Sudarmonowati E. 2017. Technological innovation in the protection of beta carotene on MOCAF production which is rich in beta carotene. Nusantara Bioscience 9: 6-11. The conversion process of high beta carotene cassava into high beta carotene MOCAF might cause a high loss of beta carotene contents during the process of flour making. This research aims to develop the most appropriate technology to protect the loss of beta carotene contents during MOCAF process. Preservation methods used consisting of four starter treatment combinations of Bimo-CF and sodium metabisulfite (Na 2 S 2 O 5 ) on Adira 1 variety, namely (i) 0.5 g starter/L volume of soaking solution and 0.3% of Na 2 S 2 O 5 , (ii) 0.5 g starter and 0.15% of Na 2 S 2 O 5 , (iii) 1 g starter and 0.3% of Na 2 S 2 O 5 , and (iv) 1 g starter and 0.15% of Na 2 S 2 O 5 . The results showed that Na 2 S 2 O 5 can reduce the loss of beta carotene contents during the process of flour making. Of the initial concentration of beta carotene in tubers (3.5 mg/kg), the loss of beta carotene on MOCAF control without preservation process with Na 2 S 2 O 5 reached 68.8% (1.09 mg/kg). With Na 2 S 2 O 5 application, the loss of beta carotene could be reduced to 24.3% (2.65 mg/kg). The fermentation process with Bimo-CF starter increased the protein content of MOCAF two times from the control. The best and safest protection of beta carotene for food was achieved from preservation method using the combination of 1 g/L volume of soaking solution starter and 0.15% of Na 2 S 2 O 5 which produced residue of Na 2 S 2 O 5 to 53.07 mg/kg, which means lower than the maximum residue allowed in a flour product (70 mg/kg). The MOCAF yield obtained from each combination is quite high in range of 32-38%. The highest MOCAF yield is obtained from the protection method using combination of 1 g/L volume of starter soaking solution with Na 2 S 2 O 5 0% and 0.15%.

Treacher Collins syndrome (TCS, OMIM: 154500) is a rare congenital craniofacial disorder that is caused by variants in the genes TCOF1, POLR1D, POLR1C, and POLR1B. Studies on the association between phenotypic variability and their relative variants are very limited. This systematic review summarized the 53 literatures from PubMed and Scopus to explore the potential TCS genotype-phenotype correlations with statistical analysis. Studies reporting both complete molecular genetics and clinical data were included. We identified that the molecular anomaly within TCOF1 (88.71%) accounted for most TCS cases. The only true hot spot for TCOF1 was detected in exon 24, with recurrent c.4369_4373delAAGAA variant is identified.While the hot spot for POLR1D, POLR1C, and POLR1B were identified in exons 3, 8, and 15, respectively. Our result suggested that the higher severity level was likely to be observed in Asian patients harboring TCOF1 variants rather than POLR1. Moreover, common 5-bp deletions tended to have a higher severity degree in comparison to any variants within exon 24 of TCOF1. In summary, this report suggested the relationship between genetic and clinical data in TCS. Our findings could be used as a reference for clinical diagnosis and further biological studies.

Background: This study aimed to evaluate the growth ability of cassava mini stem cuttings with different node number and a variety of stem cutting shapes and their correlation with starch content in the stems at initial growth stages. Methods. In this study, the viability of cassava stem cuttings was identified in two type experiments i.e. mini-stem cuttings consisting 1 and 2 nodes and shape variation of single node mini-stem cutting. Parameters observed were shoots emergence period, number of sprouting cuttings, shoots number  of individual stem cuttings, shoots height and number of leaves. In addition, starch histochemical test was  also carried out on stems of young shoots and initial stem cuttings using Lugol’s solution. Results. Both cassava stem cuttings consisting of 1 and 2 buds indicated the same survival rate of 100%. 1 bud stem cuttings with different shapes showed different survival rate, i.e. 60-80% for semicircular and fully circular cuttings and 30-40% for box shape cuttings. The difference in survival rate with different stem size is probably related to the availability of the amount of starch to support shoots growth. Observations at week 3 after planting generally showed that the stem cuttings with 2 buds were higher than those of stem with 1 bud. Conclusion: There were differences in the scores on the starch content test qualitatively with Lugol staining, in various parts of the plant originating from 1 bud and 2 bud cuttings which may indicate a breakdown of starch during shoot development.

Background:Mycobacterium tuberculosis Complex (MTBC) is a group of Mycobacterium that causes tuberculosis (TB). TB is an infectious disease that remains a global health problem. Indonesia is one of the five countries in the world where TB is the most prevalent and became the country with tle second largest rate of TB in 2014 and 2015. MTBC has high pathogenicity that can cause infections in animals and humans. The most common route of transmission is via airborne droplet nuclei and contact with animals or humans infected with TB. MTBC has many virulence factors. One of these factors is EccB5 that is encoded by eccB5 gene. EccB5 is a transmembrane protein-conserved membrane protein and could play a role in inducing damage in host cells, macrophage infection, and may correlate with active disease. The characterization of eccB5 gene needs to be studied to determine the nucleotide sequences, which may be associated with active disease. The aim of this research was to analyze the nuclotide sequences of eccB5 gene of MTBC from suspected pulmonary tuberculosis patients, SNPs of eccB5 gene and possible correlation with the disease, especially in Indonesia.Materials and Methods:Samples were collected from the Tuberculosis Laboratory, Clinical Microbiology of Dr. Soetomo Hospital Surabaya Indonesia. DNA extraction used boiling extraction method and continued nucleic acid amplification using PCR techniques. Primer pairs used eccB5 SK.. The positivity of DNA specific revealed amplicon in 1592 bp. PCR product was sequenced by 1st Base (First BASE Laboratories Sdn Bhd, Selangor, Malaysia). The sequence analysis used Genetyx-Win version 10.0 (Genetyx Corporation, Tokyo, Japan).Results:Total isolates of Mycobacterium spp. were 28 and those that showed positive MTBC were 24 isolates and 4 nontuberculosis mycobacteria (NTM) using immunochromatographic test (ICT). The amount of homology from MTBC using blast NCBI was 99%-100%. Two SNPs were found in position c.1277 which revealed replacement of amino acid in 426 of codon position.Conclusion:The sequence of eccB5 gene of MTBC showed high significant homology, while proposed non-synoymous single nucleotide polymorphisms (nsSNP) may associated with clinical outcomes.

Arginine decarboxylase (ADC) is a key enzyme responsible for polyamines biosynthesis and has been shown to increase resistance to biotic and abiotic stress. Cassava (Manihot esculenta Crantz.) is able to grow and produce storage roots well on marginal land. The purpose of this study was to optimize annealing temperature of primers in PCR reaction to amplify candidate cassava ADC gene fragments. Annealing temperature is a crucial factor in PCR reaction affecting product (gene fragments) specificity. Four pairs of primers; MeADC1, MeADC2, MeADC3, andMeADC4, were designed using degenerate method from several plants species such as Jatropa curcas (Acc XM_022220421), Populus trichocarpa (Acc XM_002306105.2), Capsicum annuum cv Nockwang (Acc KC160547.1) and Lycopersicon esculentum (Acc L16582.1). All primer pairs successfully amplified DNA fragments from local cassava genotypes (Maluku Tenggara/Malra) including Malra012 and Malra016. The MeADC1 primer amplified DNA fragment with less than 1,000 base pairs (bp) at annealing temperature of 46°C, 47°C and 48°C. However, analysis of PCR product sequencing results using NCBI BLAST method showed that the amplified DNA fragment encodes for ribosomal protein S3 of Oryza minuta (Acc YP_009242005.1).

Mycobacterium tuberculosis (M. tuberculosis) is the causative agent of tuberculosis in human. One of the major M. tuberculosis virulence factors is early secretory antigenic target of 6-kDa (ESAT-6), and EccB 5 protein encoded by eccB 5 is one of its components. EccB 5 protein is a transmembrane protein in ESX-5 system. The aim of this study is to explore the characteristics of wild-type EccB 5 and its mutant form N426I. We expressed the EccB 5 protein by cloning the mutant and wild-type eccB 5 gene in Escherichia coli (E. coli). We compared the protein structure of wild type and mutant form of EccB 5 and found changes in structure around Asn426 (loop structure) in wild type and around Ile426 (b-strand) in the mutant. The truncated recombinant protein of EccB 5 was successfully cloned and expressed using plasmid pCold I in E. coli DH5a and E. coli strain Rosetta-gami B (DE3) and purified as a 38.6 kDa protein by using the affinity column. There was no detectable adenosine triphosphatase activity in truncated forms of EccB 5 and its mutant. In conclusion, our study reveals successful cloning and protein expression of truncated form of eccB5 gene of M. tuberculosis. EccB 5 protein in ESX-5 system may be an important membrane component involved in the transport machinery of type VII secretion system, which is essential for growth and virulence.

Cassava (Manihot esculenta Crantz.) is a carbohydrate sources containing a limited amount of micronutrients, but some genotypes contain β-carotene as the precursor of vitamin A in the storage roots and leaves. Improvement of β-caroteneand minerals such as Fe / Zn content of cassava’s nutrition is mostly through by biofortification program. The storage root of β-carotene recognized by a yellow or yellowish color while the apical shoots with red to purplish. β-carotenein carotenoid biosynthetic pathway is an expression of the phytoene synthase (PSY) gene. The MePSY2 gene, one of the three MePSY family is the key gene to characterize carotenoids related gene in cassava. In this study, sequencing of the two cassava fulllenght PSY genomic DNA was carried out in conserved areas in the PSY gene region (PSY1 and PSY2) from the DNA of the cassava leaves. Adira1, Carvita25 and Ubi Kuning are yellow root storage genotypes (K1, K2 and K3) while Adira4 and Menti are white root storage genotypes (P1 and P2). Carvita25 is induced somaclonal variant of the Adira4 genotype. Contiq and consensus of nucleotide base sequences from the five cassava genotypes and CM3306-4 cultivars (acc GU111715.1) as references were analysed using the lasergene DNASTAR sequence analysis program. The results of the alignment of the base sequence constituent of the MePSY2 gene showed that the PSY2 gene with amplified genome length was 2,380 base pairs (bp) consisting of 1,140 bp exon region and 1,240 bp intron region. In the conserved coding region, there was a difference of one nucleotide base, that is, base C in two white tuber cassava genotypes replaced with A in three yellow tuber cassava genotypes in the 1.485 base (C1.485A). The SNP converts the amino acid (aa) alanine (A) to aspartic acid (D) at the 191th (A191D). Single Nucleotide polymorphism in conserved coding region can be used further as carotenoid marker for plant breeding of yellow root cassava. Keywords: β carotene, PSY gene, polymorphic gene SNP, yellow root cassava.